Table 3.

Frequency of HCMV reactivation in primitive hematopoietic subpopulations



Frequency of infectious centers*


Lysate control
Reactivation
Population phenotype
Replicates
Average
SD
Replicates
Average
SD
Fold increase§
P
CD34+         
  2.1 × 10-5  1.9 × 10-5  —   1.1 × 10-4  1.3 × 10-4  —   6.8   —  
  1.6 × 10-5    1.4 × 10-4     
CD34+/CD38-         
  2.4 × 10-5    1.0 × 10-4     
  ≤ 1.0 × 10-5  2.3 × 10-5  —   1.1 × 10-4  1.2 × 10-4  2.1 × 10-5  5.2   .002  
  3.4 × 10-5    1.4 × 10-4     
CD34+/CD38+         
  1.0 × 10-5    ≤ 1.0 × 10-5     
  ≤ 1.0 × 10-5  1.0 × 10-5  —   ≤ 1.0 × 10-5  1.0 × 10-5  —   1.0   —  
  ≤ 1.0 × 10-5    ≤ 1.0 × 10-5     
CD34+/c-kit+         
  ≤ 1.0 × 10-5    1.0 × 10-5     
  ≤ 1.0 × 10-5  1.0 × 1.0-5  —   ≤ 1.0 × 10-5  1.0 × 10-5  —   1.0   —  
  ≤ 1.0 × 10-5    1.0 × 10-5     
CD34+/c-kit-         
  1.2 × 10-5    2.6 × 10-5     
  1.0 × 10-5  1.1 × 10-5  1.2 × 10-6  1.0 × 10-5  1.6 × 10-5  8.7 × 10-6  1.6   .338  
  1.0 × 10-5    1.2 × 10-5     
CD34+/CD38-/lin-/Thy-1+         

 
1.7 × 10-4
 

 

 
3.4 × 10-4
 

 

 
2.0
 

 


Frequency of infectious centers*


Lysate control
Reactivation
Population phenotype
Replicates
Average
SD
Replicates
Average
SD
Fold increase§
P
CD34+         
  2.1 × 10-5  1.9 × 10-5  —   1.1 × 10-4  1.3 × 10-4  —   6.8   —  
  1.6 × 10-5    1.4 × 10-4     
CD34+/CD38-         
  2.4 × 10-5    1.0 × 10-4     
  ≤ 1.0 × 10-5  2.3 × 10-5  —   1.1 × 10-4  1.2 × 10-4  2.1 × 10-5  5.2   .002  
  3.4 × 10-5    1.4 × 10-4     
CD34+/CD38+         
  1.0 × 10-5    ≤ 1.0 × 10-5     
  ≤ 1.0 × 10-5  1.0 × 10-5  —   ≤ 1.0 × 10-5  1.0 × 10-5  —   1.0   —  
  ≤ 1.0 × 10-5    ≤ 1.0 × 10-5     
CD34+/c-kit+         
  ≤ 1.0 × 10-5    1.0 × 10-5     
  ≤ 1.0 × 10-5  1.0 × 1.0-5  —   ≤ 1.0 × 10-5  1.0 × 10-5  —   1.0   —  
  ≤ 1.0 × 10-5    1.0 × 10-5     
CD34+/c-kit-         
  1.2 × 10-5    2.6 × 10-5     
  1.0 × 10-5  1.1 × 10-5  1.2 × 10-6  1.0 × 10-5  1.6 × 10-5  8.7 × 10-6  1.6   .338  
  1.0 × 10-5    1.2 × 10-5     
CD34+/CD38-/lin-/Thy-1+         

 
1.7 × 10-4
 

 

 
3.4 × 10-4
 

 

 
2.0
 

 

— indicates not applicable.

*

Frequency of infectious centers is calculated by using a modified TCID50 assay based on a logistic model to allow precise extrapolation of the frequency of infectious center formation for experiments in which at least 2 wells were positive. The frequency is expressed as 1 divided by the number of cells required to yield 1 infectious center following reactivation as described in “Materials and methods.” In the event only 1 well was positive, the frequency was estimated to be 1 in 96 000 cells or 1.0 × 10-5.

A parallel series of limiting dilutions were plated following cell lysis to distinguish virus formed prior to reactivation from that formed following reactivation.

Standard deviations were calculated when at least 3 replicates yielded a reactivation frequency. SDs were not calculated for experiments in which the frequencies were estimated because of no wells scoring positively.

§

Fold increase in the frequency of reactivation compared with the lysate control.

By using the Student ttest, the probability (P value) that differences between the 2 averages were significant (P < .05) was calculated for experiments with 3 replicates.

When no wells were positive for reactivation, it was assumed that the frequency of infectious centers would be 1 or less in 25 wells or less than 1 in 96 000 cells. In this case, a value of ≤ 1.0 × 10-5 (1 in 96 000 cells) was used to estimate the upper limit of reactivation.

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